花粉管通道法验证花生Oleosin基因启动子的特异性表达

油体蛋白（Oleosin）是一类覆盖在油体表面且仅在种子中特异表达的低分子质量蛋白，能维护油体的稳定性。本研究利用花生Oleosin基因的启动子替换了pCAMBIA1301的35S启动子，构建了重组表达载体pCAMBIA1301-Oleosin-pro，利用花粉管通道法导入花生。对T0代转基因植株进行了PCR鉴定和GUS组织化学染色，结果显示：50株转基因植株有9株能扩增出目的条带，阳性率为18%，9株阳性转基因植株中有5株的子叶能被GUS染色。对GUS染色成阳性的转基因植株的子叶进行切片观察，结果显示Oleosin基因启动子驱动的GUS基因在油体蛋白中表达。

Identifition of Specific Expression of Promoter of Oleosin Gene via Pollen-tube Pathway Method

Oleosins are a kind of low molecular weight proteins that attached specifically to oil bodies, which can maintain the stability of the oil bodies. In this experiment, the recombinant expression vector pCAMBIA1301-Oleosin-pro was constructed using the promoter of Oleosin gene from peanut and transformed into peanut via pollen-tube pathway method. Transgenic plants of T0 generation were analyzed by PCR and GUS staining of tissues, the results showed that an objective band was amplified in 9 of 50 transgenic plants with the positive rate of 18%, and GUS activity was detected only in cotyledons of 5 transgenic plants. Cotyledons of positive transgenic plants identified by GUS staining were observed by microsection, which revealed that the GUS gene drived by the promoter of the Oleosin gene was expressed in the oil bodies.